摘要
目的:探索血管紧张素Ⅱ(Ang Ⅱ)引发微血管内皮通透性增加的具体机制。方法:大鼠主动脉内皮细胞原代培养鉴定,取4-7代细胞用于实验,将细胞分为对照组、Ang Ⅱ组、Ang Ⅱ+缬沙坦组。采用实时荧光定量聚合酶链反应(RT-qPCR)、蛋白质印迹(Western blot)法检测细胞中紧密连接(ZO-1)、血管内皮钙粘连蛋白(VE-cadherin)的表达,免疫荧光染色检测细胞中ZO-1蛋白的分布。通过向内皮细胞转染过表达VE-cadherin质粒,检测过表达VE-cadherin对Ang Ⅱ诱导的ZO-1蛋白表达和蛋白分布改变的影响。结果:相比于对照组,Ang II组内皮细胞ZO-1和VE-cadherin在mRNA和蛋白水平的表达均显著减少(P<0.01),且ZO-1蛋白在细胞周边分布的趋势显著降低。在Ang Ⅱ干预的内皮细胞中过表达VE-cadherin后,内皮细胞ZO-1蛋白表达增加(P<0.01),细胞周边分布趋势显著增强。结论:Ang Ⅱ通过下调VE-cadherin抑制血管内皮细胞中ZO-1蛋白的表达,从而破坏内皮细胞间的紧密连接,这可能是Ang II引发微血管通透性增加的具体机制。
Objective:To explore the mechanism by which angiotensin(Ang Ⅱ)disrupts microvascular permeability.Methods:The primary culture and identification of rat endothelial cells were conducted,and cells in passage 4-7 used for the experiment were divided as the control,Ang Ⅱ and Ang Ⅱ+valsartan groups;real-time quantitative polymerase chain reaction(RT-qPCR)and Western blot were used to test the expression of zonula occludens-1(ZO-1)and vascular endothelial(VE)-cadherin in the cells;the distribution of ZO-1 protein was detected by immunofluorescence assay.After transfection of VE-cadherin into endothelial cells,the cells were divided as the control,Ang Ⅱ and Ang Ⅱ+VE-cadherin groups.The expression and distribution of ZO-1 protein were also detected.Results:Compared with the control group,the expression of ZO-1 and VE-cadherin in Ang Ⅱ group was significantly decreased in mRNA level and protein level(both P<0.01).Additionally,the peripheral positioning of ZO-1 protein in cell membrane was remarkably depressed.However,over-expression of VEcadherin by transfecting over-expression plasmid could significantly reverse the expression and distribution of ZO-1 in Ang II-treated endothelial cells.Conclusion:Ang Ⅱ inhibited the expression of ZO-1 protein in vascular endothelial cells by down-regulating VE-cadherin,thus destroying the tight junction between endothelial cells,which may be the possible mechanism by which Ang Ⅱ disrupts microvascular permeability.
作者
沈红枫
查渭
金志江
夏海江
刘龙斌
SHEN Hongfeng;CHA Wei;JIN Zhijiang;XIA Haijiang;LIU Longbin(Department of Cardiology,the Affiliated Hospital of Shaoxing University,Shaoxing Municipal Hospital,Shaoxing 312000,China)
出处
《温州医科大学学报》
CAS
2023年第9期729-733,共5页
Journal of Wenzhou Medical University
基金
绍兴市科技计划项目(2018C30021)
浙江省医药卫生科技计划项目(2016RCA027)。
